Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies
Identifieur interne : 002190 ( Ncbi/Merge ); précédent : 002189; suivant : 002191Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies
Auteurs : Chia Chiu Lim [Malaisie] ; Patrick C. Y. Woo [République populaire de Chine] ; Theam Soon Lim [Malaisie]Source :
- Scientific Reports [ 2045-2322 ] ; 2019.
Abstract
Antibody phage display has been pivotal in the quest to generate human monoclonal antibodies for biomedical and research applications. Target antigen preparation is a main bottleneck associated with the panning process. This includes production complexity, downstream purification, quality and yield. In many instances, purified antigens are preferred for panning but this may not be possible for certain difficult target antigens. Here, we describe an improved procedure of affinity selection against crude or non-purified antigen by saturation of non-binders with blocking agents to promote positive binder enrichment termed as
Url:
DOI: 10.1038/s41598-019-42628-6
PubMed: 30988390
PubMed Central: 6465254
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PMC:6465254Le document en format XML
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies</title>
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<front><div type="abstract" xml:lang="en"><p id="Par1">Antibody phage display has been pivotal in the quest to generate human monoclonal antibodies for biomedical and research applications. Target antigen preparation is a main bottleneck associated with the panning process. This includes production complexity, downstream purification, quality and yield. In many instances, purified antigens are preferred for panning but this may not be possible for certain difficult target antigens. Here, we describe an improved procedure of affinity selection against crude or non-purified antigen by saturation of non-binders with blocking agents to promote positive binder enrichment termed as <italic>Yin-Yang</italic>
panning. A naïve human scFv library with kappa light chain repertoire with a library size of 10<sup>9</sup>
was developed. The improved <italic>Yin-Yang</italic>
biopanning process was able to enrich monoclonal antibodies specific to the MERS-CoV nucleoprotein. Three unique monoclonal antibodies were isolated in the process. The <italic>Yin-Yang</italic>
biopanning method highlights the possibility of utilizing crude antigens for the isolation of monoclonal antibodies by phage display.</p>
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<double pmid="30988390"><pmc><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies</title>
<author><name sortKey="Lim, Chia Chiu" sort="Lim, Chia Chiu" uniqKey="Lim C" first="Chia Chiu" last="Lim">Chia Chiu Lim</name>
<affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 2294 3534</institution-id>
<institution-id institution-id-type="GRID">grid.11875.3a</institution-id>
<institution>Institute for Research in Molecular Medicine,</institution>
<institution>Universiti Sains Malaysia,</institution>
</institution-wrap>
11800 Penang, Malaysia</nlm:aff>
<country xml:lang="fr">Malaisie</country>
<wicri:regionArea>11800 Penang</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Woo, Patrick C Y" sort="Woo, Patrick C Y" uniqKey="Woo P" first="Patrick C. Y." last="Woo">Patrick C. Y. Woo</name>
<affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="ISNI">0000000121742757</institution-id>
<institution-id institution-id-type="GRID">grid.194645.b</institution-id>
<institution>Department of Microbiology,</institution>
<institution>The University of Hong Kong,</institution>
</institution-wrap>
Hong Kong SAR, People’s Republic of China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>Hong Kong SAR</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Lim, Theam Soon" sort="Lim, Theam Soon" uniqKey="Lim T" first="Theam Soon" last="Lim">Theam Soon Lim</name>
<affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 2294 3534</institution-id>
<institution-id institution-id-type="GRID">grid.11875.3a</institution-id>
<institution>Institute for Research in Molecular Medicine,</institution>
<institution>Universiti Sains Malaysia,</institution>
</institution-wrap>
11800 Penang, Malaysia</nlm:aff>
<country xml:lang="fr">Malaisie</country>
<wicri:regionArea>11800 Penang</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="Aff3"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 2294 3534</institution-id>
<institution-id institution-id-type="GRID">grid.11875.3a</institution-id>
<institution>Analytical Biochemistry Research Centre,</institution>
<institution>Universiti Sains Malaysia,</institution>
</institution-wrap>
11800 Penang, Malaysia</nlm:aff>
<country xml:lang="fr">Malaisie</country>
<wicri:regionArea>11800 Penang</wicri:regionArea>
</affiliation>
</author>
</titleStmt>
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<idno type="pmid">30988390</idno>
<idno type="pmc">6465254</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6465254</idno>
<idno type="RBID">PMC:6465254</idno>
<idno type="doi">10.1038/s41598-019-42628-6</idno>
<date when="2019">2019</date>
<idno type="wicri:Area/Pmc/Corpus">000458</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000458</idno>
<idno type="wicri:Area/Pmc/Curation">000458</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000458</idno>
<idno type="wicri:Area/Pmc/Checkpoint">000343</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000343</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies</title>
<author><name sortKey="Lim, Chia Chiu" sort="Lim, Chia Chiu" uniqKey="Lim C" first="Chia Chiu" last="Lim">Chia Chiu Lim</name>
<affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 2294 3534</institution-id>
<institution-id institution-id-type="GRID">grid.11875.3a</institution-id>
<institution>Institute for Research in Molecular Medicine,</institution>
<institution>Universiti Sains Malaysia,</institution>
</institution-wrap>
11800 Penang, Malaysia</nlm:aff>
<country xml:lang="fr">Malaisie</country>
<wicri:regionArea>11800 Penang</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Woo, Patrick C Y" sort="Woo, Patrick C Y" uniqKey="Woo P" first="Patrick C. Y." last="Woo">Patrick C. Y. Woo</name>
<affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="ISNI">0000000121742757</institution-id>
<institution-id institution-id-type="GRID">grid.194645.b</institution-id>
<institution>Department of Microbiology,</institution>
<institution>The University of Hong Kong,</institution>
</institution-wrap>
Hong Kong SAR, People’s Republic of China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>Hong Kong SAR</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Lim, Theam Soon" sort="Lim, Theam Soon" uniqKey="Lim T" first="Theam Soon" last="Lim">Theam Soon Lim</name>
<affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 2294 3534</institution-id>
<institution-id institution-id-type="GRID">grid.11875.3a</institution-id>
<institution>Institute for Research in Molecular Medicine,</institution>
<institution>Universiti Sains Malaysia,</institution>
</institution-wrap>
11800 Penang, Malaysia</nlm:aff>
<country xml:lang="fr">Malaisie</country>
<wicri:regionArea>11800 Penang</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="Aff3"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 2294 3534</institution-id>
<institution-id institution-id-type="GRID">grid.11875.3a</institution-id>
<institution>Analytical Biochemistry Research Centre,</institution>
<institution>Universiti Sains Malaysia,</institution>
</institution-wrap>
11800 Penang, Malaysia</nlm:aff>
<country xml:lang="fr">Malaisie</country>
<wicri:regionArea>11800 Penang</wicri:regionArea>
</affiliation>
</author>
</analytic>
<series><title level="j">Scientific Reports</title>
<idno type="eISSN">2045-2322</idno>
<imprint><date when="2019">2019</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
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</teiHeader>
<front><div type="abstract" xml:lang="en"><p id="Par1">Antibody phage display has been pivotal in the quest to generate human monoclonal antibodies for biomedical and research applications. Target antigen preparation is a main bottleneck associated with the panning process. This includes production complexity, downstream purification, quality and yield. In many instances, purified antigens are preferred for panning but this may not be possible for certain difficult target antigens. Here, we describe an improved procedure of affinity selection against crude or non-purified antigen by saturation of non-binders with blocking agents to promote positive binder enrichment termed as <italic>Yin-Yang</italic>
panning. A naïve human scFv library with kappa light chain repertoire with a library size of 10<sup>9</sup>
was developed. The improved <italic>Yin-Yang</italic>
biopanning process was able to enrich monoclonal antibodies specific to the MERS-CoV nucleoprotein. Three unique monoclonal antibodies were isolated in the process. The <italic>Yin-Yang</italic>
biopanning method highlights the possibility of utilizing crude antigens for the isolation of monoclonal antibodies by phage display.</p>
</div>
</front>
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<pubmed><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies.</title>
<author><name sortKey="Lim, Chia Chiu" sort="Lim, Chia Chiu" uniqKey="Lim C" first="Chia Chiu" last="Lim">Chia Chiu Lim</name>
<affiliation wicri:level="1"><nlm:affiliation>Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800, Penang, Malaysia.</nlm:affiliation>
<country xml:lang="fr">Malaisie</country>
<wicri:regionArea>Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800, Penang</wicri:regionArea>
<wicri:noRegion>Penang</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Woo, Patrick C Y" sort="Woo, Patrick C Y" uniqKey="Woo P" first="Patrick C Y" last="Woo">Patrick C Y. Woo</name>
<affiliation wicri:level="1"><nlm:affiliation>Department of Microbiology, The University of Hong Kong, Hong Kong SAR, People's Republic of China.</nlm:affiliation>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>Department of Microbiology, The University of Hong Kong, Hong Kong SAR</wicri:regionArea>
<wicri:noRegion>Hong Kong SAR</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Lim, Theam Soon" sort="Lim, Theam Soon" uniqKey="Lim T" first="Theam Soon" last="Lim">Theam Soon Lim</name>
<affiliation wicri:level="1"><nlm:affiliation>Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800, Penang, Malaysia. theamsoon@usm.my.</nlm:affiliation>
<country xml:lang="fr">Malaisie</country>
<wicri:regionArea>Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800, Penang</wicri:regionArea>
<wicri:noRegion>Penang</wicri:noRegion>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PubMed</idno>
<date when="2019">2019</date>
<idno type="RBID">pubmed:30988390</idno>
<idno type="pmid">30988390</idno>
<idno type="doi">10.1038/s41598-019-42628-6</idno>
<idno type="wicri:Area/PubMed/Corpus">000567</idno>
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<idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">000604</idno>
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<sourceDesc><biblStruct><analytic><title xml:lang="en">Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies.</title>
<author><name sortKey="Lim, Chia Chiu" sort="Lim, Chia Chiu" uniqKey="Lim C" first="Chia Chiu" last="Lim">Chia Chiu Lim</name>
<affiliation wicri:level="1"><nlm:affiliation>Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800, Penang, Malaysia.</nlm:affiliation>
<country xml:lang="fr">Malaisie</country>
<wicri:regionArea>Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800, Penang</wicri:regionArea>
<wicri:noRegion>Penang</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Woo, Patrick C Y" sort="Woo, Patrick C Y" uniqKey="Woo P" first="Patrick C Y" last="Woo">Patrick C Y. Woo</name>
<affiliation wicri:level="1"><nlm:affiliation>Department of Microbiology, The University of Hong Kong, Hong Kong SAR, People's Republic of China.</nlm:affiliation>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>Department of Microbiology, The University of Hong Kong, Hong Kong SAR</wicri:regionArea>
<wicri:noRegion>Hong Kong SAR</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Lim, Theam Soon" sort="Lim, Theam Soon" uniqKey="Lim T" first="Theam Soon" last="Lim">Theam Soon Lim</name>
<affiliation wicri:level="1"><nlm:affiliation>Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800, Penang, Malaysia. theamsoon@usm.my.</nlm:affiliation>
<country xml:lang="fr">Malaisie</country>
<wicri:regionArea>Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800, Penang</wicri:regionArea>
<wicri:noRegion>Penang</wicri:noRegion>
</affiliation>
</author>
</analytic>
<series><title level="j">Scientific reports</title>
<idno type="eISSN">2045-2322</idno>
<imprint><date when="2019" type="published">2019</date>
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<front><div type="abstract" xml:lang="en">Antibody phage display has been pivotal in the quest to generate human monoclonal antibodies for biomedical and research applications. Target antigen preparation is a main bottleneck associated with the panning process. This includes production complexity, downstream purification, quality and yield. In many instances, purified antigens are preferred for panning but this may not be possible for certain difficult target antigens. Here, we describe an improved procedure of affinity selection against crude or non-purified antigen by saturation of non-binders with blocking agents to promote positive binder enrichment termed as Yin-Yang panning. A naïve human scFv library with kappa light chain repertoire with a library size of 10<sup>9</sup>
was developed. The improved Yin-Yang biopanning process was able to enrich monoclonal antibodies specific to the MERS-CoV nucleoprotein. Three unique monoclonal antibodies were isolated in the process. The Yin-Yang biopanning method highlights the possibility of utilizing crude antigens for the isolation of monoclonal antibodies by phage display.</div>
</front>
</TEI>
</pubmed>
</double>
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